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Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus

Porcine deltacoronavirus (PDCoV) and porcine sapelovirus (PSV) are two viruses that can cause diarrhoea in pigs and bring great economic loss to the pig industry. In this research, a duplex real-time quantitative polymerase chain reaction (qPCR) assay based on SYBR Green I was developed to simultane...

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Autores principales: Lu, Si-Jia, Ma, Meng-Yao, Yan, Xiao-Guang, Zhao, Fu-Jie, Hu, Wen-Yang, Ding, Qing-Wen, Ren, Hao-Jie, Xiang, Yu-Qiang, Zheng, Lan-Lan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Czech Academy of Agricultural Sciences 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10581527/
https://www.ncbi.nlm.nih.gov/pubmed/37981902
http://dx.doi.org/10.17221/79/2022-VETMED
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author Lu, Si-Jia
Ma, Meng-Yao
Yan, Xiao-Guang
Zhao, Fu-Jie
Hu, Wen-Yang
Ding, Qing-Wen
Ren, Hao-Jie
Xiang, Yu-Qiang
Zheng, Lan-Lan
author_facet Lu, Si-Jia
Ma, Meng-Yao
Yan, Xiao-Guang
Zhao, Fu-Jie
Hu, Wen-Yang
Ding, Qing-Wen
Ren, Hao-Jie
Xiang, Yu-Qiang
Zheng, Lan-Lan
author_sort Lu, Si-Jia
collection PubMed
description Porcine deltacoronavirus (PDCoV) and porcine sapelovirus (PSV) are two viruses that can cause diarrhoea in pigs and bring great economic loss to the pig industry. In this research, a duplex real-time quantitative polymerase chain reaction (qPCR) assay based on SYBR Green I was developed to simultaneously detect PDCoV and PSV. No specific melting peaks were found in other porcine diarrhoea-associated viruses, indicating that the method developed in this study had good specificity. The detection limits of PDCoV and PSV were 1.0 × 10(1) copies μl(–1) and 1.0 × 10(2) copies μl(–1), respectively. The duplex real-time qPCR assay tested two hundred and three (203) intestinal and faecal samples collected from diarrhoeal and asymptomatic pigs. The positive rates of PDCoV and PSV were 20.2% and 23.2%, respectively. The co-infection rate of PDCoV and PSV was 13.8%. To evaluate the accuracy of the developed method, conventional PCR and singular TaqMan real-time qPCR assays for PDCoV/PSV were also used to detect the samples. The results showed that the duplex real-time qPCR assay was consistent with the singular assays, but its sensitivity was higher than conventional PCR methods. This duplex real-time qPCR assay provides a rapid, sensitive and reliable method in a clinic to simultaneously detect PDCoV and PSV.
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spelling pubmed-105815272023-11-17 Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus Lu, Si-Jia Ma, Meng-Yao Yan, Xiao-Guang Zhao, Fu-Jie Hu, Wen-Yang Ding, Qing-Wen Ren, Hao-Jie Xiang, Yu-Qiang Zheng, Lan-Lan Vet Med (Praha) Original Paper Porcine deltacoronavirus (PDCoV) and porcine sapelovirus (PSV) are two viruses that can cause diarrhoea in pigs and bring great economic loss to the pig industry. In this research, a duplex real-time quantitative polymerase chain reaction (qPCR) assay based on SYBR Green I was developed to simultaneously detect PDCoV and PSV. No specific melting peaks were found in other porcine diarrhoea-associated viruses, indicating that the method developed in this study had good specificity. The detection limits of PDCoV and PSV were 1.0 × 10(1) copies μl(–1) and 1.0 × 10(2) copies μl(–1), respectively. The duplex real-time qPCR assay tested two hundred and three (203) intestinal and faecal samples collected from diarrhoeal and asymptomatic pigs. The positive rates of PDCoV and PSV were 20.2% and 23.2%, respectively. The co-infection rate of PDCoV and PSV was 13.8%. To evaluate the accuracy of the developed method, conventional PCR and singular TaqMan real-time qPCR assays for PDCoV/PSV were also used to detect the samples. The results showed that the duplex real-time qPCR assay was consistent with the singular assays, but its sensitivity was higher than conventional PCR methods. This duplex real-time qPCR assay provides a rapid, sensitive and reliable method in a clinic to simultaneously detect PDCoV and PSV. Czech Academy of Agricultural Sciences 2023-03-23 /pmc/articles/PMC10581527/ /pubmed/37981902 http://dx.doi.org/10.17221/79/2022-VETMED Text en Copyright: © 2023 Lu et al. https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International (https://creativecommons.org/licenses/by/4.0/) (CC BY-NC 4.0).
spellingShingle Original Paper
Lu, Si-Jia
Ma, Meng-Yao
Yan, Xiao-Guang
Zhao, Fu-Jie
Hu, Wen-Yang
Ding, Qing-Wen
Ren, Hao-Jie
Xiang, Yu-Qiang
Zheng, Lan-Lan
Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus
title Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus
title_full Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus
title_fullStr Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus
title_full_unstemmed Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus
title_short Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus
title_sort development and application of a low-priced duplex quantitative pcr assay based on sybr green i for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10581527/
https://www.ncbi.nlm.nih.gov/pubmed/37981902
http://dx.doi.org/10.17221/79/2022-VETMED
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